title: Molecular recognition of a model globular protein apomyoglobin by synthetic receptor cyclodextrin: effect of fluorescence modification of the protein and cavity size of the receptor in the interaction.
abstract: Labelling of proteins with some extrinsic probe is unavoidable in molecular biology research. Particularly, spectroscopic studies in the optical region require fluorescence modification of native proteins by attaching polycyclic aromatic fluoroprobe with the proteins under investigation. Our present study aims to address the consequence of the attachment of a fluoroprobe at the protein surface in the molecular recognition of the protein by selectively small model receptor. A spectroscopic study involving apomyoglobin (Apo-Mb) and cyclodextrin (CyD) of various cavity sizes as model globular protein and synthetic receptors, respectively, using steady-state and picosecond-resolved techniques, is detailed here. A study involving Förster resonance energy transfer, between intrinsic amino acid tryptophan (donor) and N, N-dimethyl naphthalene moiety of the extrinsic dansyl probes at the surface of Apo-Mb, precisely monitor changes in donor acceptor distance as a consequence of interaction of the protein with CyD having different cavity sizes (β and γ variety). Molecular modelling studies on the interaction of tryptophan and dansyl probe with β-CyD is reported here and found to be consistent with the experimental observations. In order to investigate structural aspects of the interacting protein, we have used circular dichroism spectroscopy. Temperature-dependent circular dichroism studies explore the change in the secondary structure of Apo-Mb in association with CyD, before and after fluorescence modification of the protein. Overall, the study well exemplifies approaches to protein recognition by CyD as a synthetic receptor and offers a cautionary note on the use of hydrophobic fluorescent labels for proteins in biochemical studies involving recognition of molecules.
authors.affiliation: Department of Chemical, Biological and Macromolecular Sciences, S.N. Bose National Centre for Basic Sciences, Block JD, Sector III, Salt Lake, Kolkata, 700098, India.
authors.forename: Ranajay
authors.initials: R
authors.lastname: Saha
authors.forename: Surajit
authors.initials: S
authors.lastname: Rakshit
authors.forename: Samir Kumar
authors.initials: SK
authors.lastname: Pal
mesh_terms: D000818:Animals; D001059:Apoproteins / Q000737:chemistry* / Q000378:metabolism*; D002942:Circular Dichroism; D006736:Horses; D008958:Models, Molecular; D009211:Myoglobin / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013997:Time Factors; D014364:Tryptophan / Q000737:chemistry; D047392:beta-Cyclodextrins / Q000737:chemistry* / Q000378:metabolism*; D047408:gamma-Cyclodextrins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D001059:Apoproteins; D009211:Myoglobin; C010857:apomyoglobin; D047392:beta-Cyclodextrins; D047408:gamma-Cyclodextrins; D014364:Tryptophan; C031215:betadex
keywords: apomyoglobin; cyclodextrin; fluorescence labeling; molecular recognition; receptor cavity size